■本研究通过生长停滞特异性转录因子5(GAS5)/microRNA-21(miR-21)/Toll样受体4(TLR4)轴,探讨了新风胶囊(XFC)在佐剂性关节炎(AA)大鼠心肌损伤中的作用机制。
■大鼠注射弗氏完全佐剂建立大鼠AA模型。然后,一些模型大鼠只给生理盐水或药物,一些模型大鼠在给药前注射腺相关病毒或神经磺胺(NSA;一种焦凋亡抑制剂)。计算脚趾肿胀和关节炎指数(AI)。用苏木精-伊红染色分析滑膜和心肌组织的病理和形态学变化,透射电镜观察心肌组织的变性囊泡和超微结构变化。血清白细胞介素(IL)-1β水平,检测IL-18、IL-6和肿瘤坏死因子(TNF)-α,并在心肌组织中测量乳酸脱氢酶(LDH)的释放,伴随GAS5,miR-21,TLR4,核因子-kB(NF-κB)p65,核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)的检查,Caspase-1和GasderminD(GSDMD)在心肌组织中的表达。
■AA建模后,大鼠出现显著增加的脚趾肿胀和AI评分,滑膜和心肌组织损伤,高变性囊泡,血清IL-1β水平显著升高,IL-18、IL-6和TNF-α,伴随着GAS5表达的显着减少,显著增强miR-21、TLR4、NF-κBp65、NLRP3、Caspase-1和GSDMD表达,显著增加心肌组织中LDH的释放。XFC治疗显著下降脚趾肿胀,AI分数,滑膜和心肌组织损伤,血清IL-1β水平,AA大鼠的IL-18、IL-6和TNF-α。此外,XFC治疗可显著升高AA大鼠心肌组织中GAS5的表达并显著降低LDH的释放和miR-21、TLR4、NF-κBp65、NLRP3、Caspase-1和GSDMD的表达。此外,GAS5过表达或NSA治疗可进一步促进XFC对AA大鼠的上述作用。
■XFC通过调节GAS5/miR-21/TLR4轴,抑制焦凋亡和促炎细胞因子分泌,减轻AA大鼠心肌损伤。
UNASSIGNED: This study probed the mechanism of action of Xinfeng Capsule (XFC) in myocardial injury in rats with adjuvant arthritis (AA) via the growth arrest-specific transcript 5 (GAS5)/microRNA-21 (miR-21)/Toll-like receptor 4 (TLR4) axis.
UNASSIGNED: Rats were injected with Freund\'s complete adjuvant to establish a rat model of AA. Then, some modeled rats were given normal saline or drugs only, and some modeled rats were injected with adeno-associated viruses or necrosulfonamide (NSA; a pyroptosis inhibitor) before drug administration. Toe swelling and arthritis index (AI) were calculated. Pathological and morphological changes in synovial and myocardial tissues were analyzed with hematoxylin-eosin staining, and pyroptotic vesicles and the ultrastructural changes of myocardial tissues were observed with transmission electron microscopy. The serum levels of interleukin (IL)-1β, IL-18, IL-6, and tumor necrosis factor (TNF)-α were detected, and lactate dehydrogenase (LDH) release was measured in myocardial tissues, accompanied by the examination of GAS5, miR-21, TLR4, nuclear factor-kB (NF-κB) p65, nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), Caspase-1, and Gasdermin D (GSDMD) expression in myocardial tissues.
UNASSIGNED: After AA modeling, rats presented with significantly increased toe swelling and AI scores, synovial and myocardial tissue damage, elevated pyroptotic vesicles, and markedly enhanced serum levels of IL-1β, IL-18, IL-6, and TNF-α, accompanied by significantly diminished GAS5 expression, substantially augmented miR-21, TLR4, NF-κB p65, NLRP3, Caspase-1, and GSDMD expression, greatly increased LDH release in myocardial tissues. XFC treatment significantly declined toe swelling, AI scores, synovial and myocardial tissue damage, and the serum levels of IL-1β, IL-18, IL-6, and TNF-α in AA rats. Additionally, XFC treatment markedly elevated GAS5 expression and substantially lowered LDH release and miR-21, TLR4, NF-κB p65, NLRP3, Caspase-1, and GSDMD expression in myocardial tissues of AA rats. Moreover, the above effects of XFC in AA rats were further promoted by GAS5 overexpression or NSA treatment.
UNASSIGNED: XFC alleviated myocardial injury in AA rats by regulating the GAS5/miR-21/TLR4 axis and inhibiting pyroptosis and pro-inflammatory cytokine secretion.